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Freeze-drying method for specific protein of central nervous system

A central nervous system-specific technology, applied in the field of central nervous system-specific protein freeze-drying, can solve the problem of no effective freeze-drying methods and freeze-drying reagents for improving the preservation activity of central nervous system-specific proteins, so as to improve the preservation effect, Effect of Improving Detection Sensitivity

Active Publication Date: 2021-07-09
SOPHONIX CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Although there are many freeze-drying methods and freeze-drying buffers in the prior art, there are no freeze-drying methods and freeze-drying reagents that can effectively improve the preservation activity of central nervous system-specific proteins.

Method used

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  • Freeze-drying method for specific protein of central nervous system
  • Freeze-drying method for specific protein of central nervous system
  • Freeze-drying method for specific protein of central nervous system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] A method for freeze-drying of central nervous system-specific protein (S100-β), comprising the following steps:

[0041] (1) Prepare buffer

[0042] The buffer solution includes the following components: 1.0 g of HEPES, 7 g of sodium chloride, 1 g of potassium chloride, 3 g of magnesium sulfate, 30 g of BSA, 30 g of trehalose, 10 g of glycine, 15 g of Casein, and 100 ml of glycerol.

[0043] The preparation method of described buffer solution, comprises the following steps:

[0044] Weigh HEPES, NaCl, KCl, MgSO 4 , trehalose, bovine serum albumin, glycine, casein, and glycerin were added to purified water and stirred until completely dissolved, adjusted to pH 7.1, and fixed to 1000ml; filtered with a filter membrane with a pore size of 0.22 μm to obtain buffer.

[0045] (2) Take the S100-β recombinant protein and dissolve it in buffer, mix it well and prepare 6 concentrations of S100-β reagent as a calibration product, the concentrations are 0ng / mL, 1ng / mL, 20ng / mL, ...

Embodiment 2

[0051] A method for freeze-drying of central nervous system-specific protein (S100-β), comprising the following steps:

[0052] (1) Prepare buffer

[0053] The buffer solution includes the following components: HEPES 10g, sodium chloride 5g, potassium chloride 5g, magnesium sulfate 10g, BSA 4.0g, trehalose 100g, glycine 4.0g, Casein 25g, glycerin 20ml.

[0054] The preparation method of described buffer solution, comprises the following steps:

[0055] Weigh HEPES, NaCl, KCl, MgSO 4 , trehalose, bovine serum albumin, glycine, casein, and glycerin were added to purified water and stirred until completely dissolved, adjusted to pH 8.2, and fixed to 1000ml; filtered with a filter membrane with a pore size of 0.22 μm to obtain buffer.

[0056] (2) Take the S100-β recombinant protein and dissolve it in buffer, mix it well and prepare 6 concentrations of S100-β reagent as a calibration product, the concentrations are 0ng / mL, 1ng / mL, 20ng / mL, 50ng / mL , 100ng / mL, 150ng / mL.

[005...

Embodiment 3

[0061] A method for freeze-drying of central nervous system-specific protein (S100-β), comprising the following steps:

[0062] (1) Prepare buffer

[0063] The buffer solution includes the following components: 3.0 g of HEPES, 9 g of sodium chloride, 0.5 g of potassium chloride, 0.1 g of magnesium sulfate, 27 g of BSA, 10 g of trehalose, 18 g of glycine, 0.5 g of Casein, and 200 ml of glycerol.

[0064] The preparation method of described buffer solution, comprises the following steps:

[0065] Weigh HEPES, NaCl, KCl, MgSO 4 , trehalose, bovine serum albumin, glycine, casein, and glycerin were added to purified water and stirred until completely dissolved, adjusted to pH 6.5, and fixed to 1000ml; filtered with a filter membrane with a pore size of 0.22 μm to obtain buffer.

[0066] (2) Take the S100-β recombinant protein and dissolve it in buffer, mix it well and prepare 6 concentrations of S100-β reagent as a calibration product, the concentrations are 0ng / mL, 1ng / mL, 20ng...

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Abstract

The invention belongs to the technical field of freeze-drying, and particularly relates to a freeze-drying method for the specific protein of a central nervous system. The freeze-drying method for the specific protein (S100-beta) of the central nervous system comprises the following steps: dissolving S100-beta in a buffer solution to prepare an S100-beta reagent, and putting the S100-beta reagent into a vacuum freeze-drying machine for freeze-drying, wherein the buffer solution is prepared from the following parts: 4-hydroxyethyl piperazine ethanesulfonic acid (HEPES), sodium chloride (NaCl), potassium chloride (KCl), magnesium sulfate (MgSO4), bovine serum albumin (BSA), trehalose, glycine, casein (Casein) and glycerol. The freeze-drying method can effectively preserve the specific protein of the central nervous system, and is beneficial to keeping the maximum activity of the specific protein of the central nervous system.

Description

technical field [0001] The invention belongs to the technical field of freeze-drying, and in particular relates to a freeze-drying method for central nervous system-specific proteins. Background technique [0002] Accurate evaluation of the severity and prognosis of central nervous system damage after traumatic brain injury (traumatic brain injury, TBI) is very important for clinical treatment. At present, there is no more accurate means to evaluate the severity of TBI, and it is difficult to judge the prognosis. Cognitive dysfunction, motor dysfunction, sensory dysfunction, emotional and emotional disorders caused by central nervous system damage seriously affect the quality of life of patients. [0003] CNS-specific protein (S100-β) is an acidic calcium-binding protein with a molecular weight of 21KD, which is mainly produced by astrocytes, forms disulfide bonds through cysteine ​​residues, and is abundant in the active form of dimers present in the central nervous syste...

Claims

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Application Information

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IPC IPC(8): C07K14/47C07K1/00G01N33/68G01N33/53
CPCC07K14/4728G01N33/68G01N33/5306G01N2333/4727
Inventor 李博飞
Owner SOPHONIX CO LTD
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